Anti-TFEB antibody [BLR070G]

4 product images

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  Immunoprecipitation - Anti-TFEB antibody [BLR070G] (ab264421)

  TFEB was immunoprecipitated from HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate (1.0 mg per IP reaction; 20% of IP loaded) with ab264421 at 20 μl/mg lysate. Western blot was performed from the immunoprecipitates using ab264421 at 1/1000 dilution.
  

  Lane 1: ab264421 IP in HeLa whole cell lysate.
  Lane 2: Control IgG IP in HeLa whole cell lysate.

  Detection: Chemiluminescence with an exposure time of 75 seconds.

  Lysates prepared using NETN lysis buffer.

  All lanes: Immunoprecipitation - Anti-TFEB antibody [BLR070G] (ab264421)

  Predicted band size: 53 kDa

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  Western blot - Anti-TFEB antibody [BLR070G] (ab264421)

  Lysates prepared using NETN lysis buffer.

  All lanes: Western blot - Anti-TFEB antibody [BLR070G] (ab264421) at 1/1000 dilution

  Lane 1: HeLa whole cell lysate at 50 µg

  Lane 2: A549 whole cell lysate at 50 µg

  Lane 3: HepG2 whole cell lysate at 50 µg

  Lane 4: Jurkat whole cell lysate at 50 µg

  Lane 5: RKO whole cell lysate at 50 µg

  Lane 6: K562 whole cell lysate at 50 µg

  Lane 7: LNcaP whole cell lysate at 50 µg

  Lane 8: MCF7 whole cell lysate at 50 µg

  Lane 9: TCMK-1 whole cell lysate at 50 µg

  Lane 10: NIH/3T3 whole cell lysate at 50 µg

  Secondary

  All lanes: HRP-conjugated goat anti-rabbit IgG

  Developed using the ECL technique.

  Predicted band size: 53 kDa

  Exposure time: 3min

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  Western blot - Anti-TFEB antibody [BLR070G] (ab264421)

  False colour image of Western blot: Anti-TFEB antibody [BLR070G] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab264421 was shown to bind specifically to TFEB. A band was observed at 60 kDa in wild-type HepG2 cell lysates with no signal observed at this size in TFEB knockout cell line Human TFEB knockout Hep G2 cell line ab277871 (knockout cell lysate ab284220). To generate this image, wild-type and TFEB knockout HepG2 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.

  All lanes: Western blot - Anti-TFEB antibody [BLR070G] (ab264421) at 1/1000 dilution

  Lane 1: Wild-type HepG2 cell lysate at 20 µg

  Lane 2: TFEB knockout HepG2 cell lysate at 20 µg

  Lane 2: Western blot - Human TFEB knockout Hep G2 cell line (Human TFEB knockout Hep G2 cell line ab277871)

  Lane 3: Raji cell lysate at 20 µg

  Lane 4: MCF7 cell lysate at 20 µg

  Performed under reducing conditions.

  Predicted band size: 53 kDa

  Observed band size: 60 kDa

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  Flow Cytometry - Anti-TFEB antibody [BLR070G] (ab264421)

  Flow cytometry overlay histogram of 4% formaldehyde fixed Daudi cells permeabilized with 90% methanol labeling human TFEB with ab264421 at 1 µl per 1 x 10^6 cells (shaded) compared with a Isotype control (unshaded). Secondary antibody details: Goat Anti-Rabbit IgG H&L (DyLight® 650) (Goat Anti-Rabbit IgG H&L (DyLight® 650) ab96886).